Mouse Retinal Organoid Growth and Maintenance in Longer-Term Culture

• DOI: 10.3389/fcell.2021.645704
• Publication Date: 2021-04-28T09:29:31Z
• Authors: Manuela Völkner; Thomas Kurth; Jana Schor; Lynn J. A. Ebner; Lara Bardtke; Cagri Kavak; Jörg Hackermüller; Mike O. Karl
• Keywords: Organoid

Using retinal organoid systems, organ-like 3D tissues, relies implicitly on their robustness. However, essential key parameters, particularly growth and longer-term culture, are still insufficiently defined. Here, we hypothesize that a previously optimized protocol for high yield of evenly-sized mouse organoids with low variability facilitates assessment such parameters. We demonstrate these reliably complete retinogenesis, can be maintained at least up to 60 days in culture. During this time, the continue mature molecular (ultra)structural level: They develop photoreceptor outer segments synapses, transiently maintain its cell composition about 5–10 after completing subsequently pathologic changes – mainly inner but also retina reactive gliosis. To test whether system provides experimental access during upon completion development, defined stimulated by activating sonic hedgehog signaling, which patients mice vivo congenital defect leads enlarged eyes. signaling activator increased epithelia length when applied not development. This experimentally supports maturation, stability, reproducibility system, potential pathology model, as well producing larger organoids. Together, our study advances understanding growth, maintenance, further optimizes future utilization.